A Report on Industrial visit

Submitted to
Department of Biotechnology
Mangalayatan UniversityAligarh
Submitted By
RAHUL KUMAR
20120366
B.Tech-BT(SEM-VIIIth)

Institute of Engineering and Technology

Mangalayatan University Beswan, Aligarh UP (INDIA)

Session: 2015-2016

CERTIFICATE
This is to certify that Mr. Rahul Kumar a student of
B.Tech-BT Final year Enrollment no. 20120366 has
successfully completed his industrial visit in "Arbro
Pharmaceuticals Limited" dated on 20th January 2016
as a part of B.Tech-BT curriculum.

Date:
Place:

Er. Pawan Sharma

Ass. Professor

ACKNOWLEDEMENT
This visit report "Arbro Pharmaceuticals Limited" is
acombined effort of intellectual person who assisted in
providing a great Knowledge in the infrastructure of this
report.
I would like to devote my first and foremost gratitude to Er.
Pawan Sharma, supervisor of Pharmaceutics department,
who took all the painstaking jobs and encouraged me whole
hearty to make this project come up with its best look.
I would not be justified if I forget to pay my due thanks to
our worthy HOD Dr. GAURAV MUDGAL who provided
me with all the necessities to compose and to make this
project facilitate in the final shape.
I would like to devote my parents who advised to me for this
report and also like to my friends who helped me in my
report completed.

About Arbro Pharmaceuticals Limited

ARBRO Analytical Division is a multi-disciplinary contract research Organization and Govt.
approve commercial test

house.

Over the last 15 years we have established ourselves as a source of consistent and prompt
service.
Arbro is a leading multidisciplinary contract research organization and testing lab in India.
Established in the year 1990 having over two decades of experience in pharmaceutical
testing, food testing,herbal testing, water testing and construction material. Working with premier
clients across industries and around the world. Arbro offers analytical method development,
method validation, stability testing, product testing, product development, chemical testing lab
and microbiology testing lab services along with preshipment inspection. Arbro has a team of
over 150 professionals spread across multiple analytical labs, backed by cutting edge technology
and world-class infrastructure.
The organizations comprehensive management and quality systems enable it to complete the
most complex projects accurately, responsively and in rigorous compliance with applicable
regulations. The laboratory is accredited by (NABL) National Accreditation Board for Testing
and Calibration (ISO/IEC 17025:2005) and ISO 9001:2008 Certified analytical laboratory that is
committed to provide the highest quality of all offered services with the lowest turn-around time.
Be it quality control testing, routine & non-routine analysis or environmental and analytical
consultancy, Arbro is the turnkey contract analysis outsourcing solution.
Arbro is recognized as a quality solution provider and partners with MNCs and leaders of
various industries to provide solutions to their analytical problems. We are respected in industry
circles as a provider of quality services with the highest level of integrity and transparency.
A loyal client base of over 10,000 satisfied customers ranging from large corporations, to
academic institutions, to small and medium businesses, to government organizations and
individuals are a testimony to our consistent performance.

Some tests performed in the lab are:

Pharmaceutical Testing

Food Testing

Water Testing

Testing of Herbal products and Nutraceuticals

Medical Device Testing

Testing of Cosmetic Products

Building and Construction Material Testing

Pre-shipment inspection and sampling of consignments

Environmental monitoring and testing of environmental samples

Atomic Absorption Spectrometer (AAS)

Atomic Absorption Spectrometer (AAS)

Atomic absorption spectroscopy is a widely used technique for trace analysis of elements

in matrices ranging from foods, waste waters, geological, pharmaceutical and biological.
The absorption occurs when a ground state atom absorbs energy in the form of light of a
specific wavelength and is elevated to an excited state. The amount of light energy
absorbed will increase as the atom population of the selected dement increases in the light

path.
ARBRO equipped with the state of art Perkin Elmer 400 atomic absorption spectrometer
which provides convinent analysis in the ppm level for major constituents .The typical
samples analysed are feed, food, water, metals, drugs and salts.

Fourier Transform Infrared Spectroscopy (FTIR)

FTIR testing lab

Infrared spectroscopy is used widely in both research and industry as a simple and rapid
technique for measurement and quality control in both organic and inorganic analysis.In
contrast to the relatively few absorption peaks observed in UV-VIS region for most

compounds, the infrared spectrum provides a rich array of absorption bands.

With the developments in interferometer designs and electronics FT-IR spectroscopy has
emerged as a technique that in a true sense provides a fingerprint of materials in a

matter of seconds.
They equipped with Jasco-410 FT-IR in our laboratory in Delhi and PerkinElmer FT-IR

in Bangalore and Baddi.

The instrument is extensively used in analysis pharmaceuticals, chemical, polymers,
foods and mineral oils in water samples.

LCMS/MS

LCMSMS
Liquid chromatography-Mass spectrometry (LC-MS) combines the physical separation
capabilities of liquid chromatography with the mass analysis capabilities of a mass spectrometer.
LC-MS is emerging as the most favored technique in major application areas including
pharmacokinetic studies and proteomics.
The Major development has been the tandem MS-MS which allows the detector to select certain
ions to fragment. With the concurrent development of HPLC now days it is possible to analyse
samples in a minute or less as compared to over 10 minutes with other detectors. At the present
growth rate the technique is slated to rank as a topmost preferred separation and analysis
technique.
ARBRO is equipped with API-300 LC-MS-MS which is operating since 2006 and a Agilent
6410 since 2010. Our LCMSMS are being put to use in following areas.

Method development and validation

Antibiotic residues in food products.

Pesticide residues in foods more than 200 compounds in various matrices

Molecular weight estimation

Impurity Profiling

Cleaning validation studies

Bio analytical-BA/BE Studies

Proteins

HPLC

HPLC
High Performance Liquid Chromatography (HPLC) is an essential analytical tool in any testing
lab as it affords separation and quantification of a variety of analytes which are thermally labile
and not volatile enough for analysis on gas chromatograph.
Arbro has 26 HPLC instruments from Waters, Jasco, Perkin Elmer and Agilent equipped with a
variety of detectors. The number of instruments is indicative of the utility of the technique in the
laboratory.
The different modes and choice of detectors permit analysis of wide range of samples, and we
have the following detectors available in our testing laboratories

UV Detector

Refractive index (RI) detector

Photo diode array detector (PDA)

Fluorescence detector (FLD)

Mass spectrometer detector

The HPLCs in testing lab are equipped with auto-samplers for 247 operation and higher
accuracy and precision. This enables us to provide reliable results and lowest turn around time
(TAT).
The large variety of stationary phases available make it one of the most versatile techniques and
it can be used for the analysis of molecules like

Small molecules

Semi polar molecules by reverse phase chromatography

Macro-molecules by Gel permeation chromatography

Polar molecules in HILIC or Cyano or Amino colums and Ion pairing chromatography

Nonpolar molecules using Silica of C1-C4 colums

HPLC is used in testing to analyze products from the following industries

Pharmaceuticals

Foods

Pesticide formulations

Water and waste water

Fine chemicals

Colors and dyes

Proteins

Bio molecules and

Environmental analysis

Following analysis on HPLC

Assay of drugs products and formulations

Impurity profiling of pharmaceuticals

Quantification of Vitamins

Estimation of Aflatoxins

Pesticide residue testing

Method development

Method validation

ICP-MS

ICP MS Lab

ICP-MS is the synergistic combination of an ICP with a mass spectrometer. ICP-MS

offers the benefits of speed of analysis along with lowest detection limits

to laboratories performing trace metal determinations.

The detection limits are at or below parts per trillion (ppt) range are achievable with ICPMS.The technique is much faster than AAS and graphite furnace AAS for determinations

of multiple elements per sample. As many as 85 elements in a sample can be analysed in

2-3 minutes.
Laboratory is equipped with Perkin Elmer DRC-e model ICP-MS along with dynamic
reaction cell which completely eliminates interferences thus providing ultra trace
detection capabilities even in difficult matrices such as waste waters, soils, sea water and

clinical samples.
For sample preparation the laboratory is equipped with a Microwave Digestion system
allowing complete and reproducible sample digestion which is critical for the elemental

analysis in complex matrices.

The facility has been needful for analysis of wide ranging samples such as Ayurvedic
products, foods and natural products such as milk, honey, drugs and drinking

GC-MS and GC-MS-MS

GCMSMS
Gas Chromatography-Mass spectroscopy (GC-MS) is a highly sensitive technique for detection
and quantification of organic molecules. The system is finding application in pharmaceuticals,
foods and environmental analysis.
ARBRO laboratory is equipped with a Perkin Elmer 5oo GC-MS. Coupling of the system with
Perkin Elmer 500 Turbomatrix GC-HS allow mass spectroscopic detection of volatile residues.

ARBRO also have an Agilent 7000 series GCMSMS which provides unparalleled sensitivity for
the detection of toxins and pesticides residues.

The GC-MS system is equipped with ECD and NPD detectors for analysis of Toxics Dioxane,
Residual, pesticides and cholesterol in agricultural samples and food samples, such milk, honey,
grapes etc.
GCMSMS system for the following

Pesticide residue analysis in food and environmental samples

Identification of unknown volatile and semi-volatile organics in pharmaceutical drug

products and substances using GC-Head space MS

Chiral analysis of amino-acid composition of protein samples

Impurity profiling in pesticides

Analysis of flavor and fragrances

Identification of unknown compounds by library matching in the NIST MS library.

Bio-analysis for organic molecules in blood, plasma, tissue and urine

Malvern Mastersizer 3000 Particle Size Analyzer

Malvern Particle Size Analyzer Mastersizer 3000

Particle size analysis are routinely required by industries as a critical control parameter for raw
materials, manufacturing processes and finished products. To fulfil this requirement of our
esteemed customers, like you, we have installed the latest; Laser Diffraction based Particle Size
Analyzer.
The System is equipped with both wet and dry mode accessories allowing the analysis of both
powders and liquid dispersion. This new system has a large linear range of measurement from 10
nanometer to 3.5 millimetres allowing particle size measurements of diverse samples ranging
from nano materials, right up to, coarse systems like pharmaceutical tablet granules.
Importance of Particle Analysis
Particle size and particle size distribution are amongst the most important physical properties of a
particulate system, as particle size has direct effect on a number of material properties, some of
these are listed below
1. Dissolution rate and bioavailability of pharmaceutical products
2. Chemical stability of reactive or hygroscopic materials
3. Physical stability of suspension and emulsions in pharmaceuticals, pigments and paints

4. Flow properties of the powder and granules in manufacturing processes

5. Efficacy of drug delivery system like inhalers
6. Reactivity of chemicals and catalyst
7. Texture and feel of food products, creams, ointments, cosmetics
8. Appearance of powder coatings and inks
9. Viscosity, especially in the case of sprays
10. Packing density and porosity in ceramic
Common Techniques Used for Particle Analysis
Depending on the particle size range and the nature of data required (size and/or shape and/or
surface characteristics) various techniques are available for the measurement of particle size.
Some of the main techniques are listed below
1. Mechanical Dry or Wet Sieving
2. Optical Microscopy
3. Sedimentation Rate using Andreasen apparatus
4. Coulter Counter, which measures particle volume
5. Dynamic Light Scattering (DLS) mostly used for the sub micrometre size range
6. Scanning Electron Microscopy (SEM)
7. Transmission Electron Microscopy (TEM)
8. Atomic Force Microscopy (AFM)
9. Automated imaging

10. Laser Diffraction the principle used in Malvern Mastersizier 3000

Malvern Mastersizer based on Laser Diffraction is the Gold Standard for Particle Analysis
Laser Diffraction in general and the Malvern Mastersizer range of particle size analyser in
particular are recognised as the industry gold standard for particle size analysis. They have
gained the reputation because of the excellent accuracy and repeatability and reproducibility
offered by the system as can be seen summarised in the table below.

Malvern Mastersizer MS3000 Performance Summary

Size range

0.01-3500m

Accuracy

Better than 1%

Repeatability

Better than 0.5%

Reproducibility

Better than 1%

Angular range

0.015-144

Red light source

Max. 5mW He-Ne, 632.8nm

Blue light source

Max. 10mW LED, 470nm

Laser diffraction is accepted and prescribed by various Pharmacopoeias, ASTM, DIN, ISO and
numerous other national and international standard methods for particle size measurements. It
finds applications in diverse industries ranging from chemicals, fine chemicals, pharmaceuticals,
food ingredients, food products, minerals, ores, and cement et cetera.
Arbro is one of the largest contact testing laboratories in India having two state-of-the-art
laboratories and a team of over 200 skilled professionals. In keeping with our policy of Offering
the most technologically advanced portfolio of services to our customers we have made this
investment to expand the scope of our services based on feedback from our customers.

Liquid Particle Counter

Liquid Particle Counter Test for Particulate Matter

The test for particulate contamination in parenteral preparations is a critical test and is a
mandatory pharmacopoeial requirement. The pharmacopoeias prescribe either a microscopic
particle count or a light obscuration liquid particle counter, to carry out this test. We
at Arbro have been offering the microscopic particle count test for many years now and are
pleased to share that we have recently installed the light obscuration of a liquid particle counter
in our laboratory, to fulfill this frequent requirement from our customers involved in the
manufacturing of parenteral preparations.

What is particulate contamination?

The pharmacopoeia defines particulate contamination as the unintentional presence in injections
and infusions, of extraneous, mobile, un-dissolved substances, other than gas bubbles. These
solids may consist of individual components or mixture of cellulose, glass, rubber cores from
vials and plastic fragments

What are potential source of particulate contamination?

Even though sterile manufacturing facilities and processes are designed and controlled to
minimise the generation of particulate matter. Still, it is possible for contamination from any of
the following sources to enter the product
1. Manufacturing environment
2. Process equipment
3. Manufacturing personal
4. The packaging components
5. Can be generated by chemical reaction or incompatibility of ingredients of the
formulation.

Why should it be controlled?

The test for particulate matter primarily deals with non-infectious particles which may be present
in an intravenous formulation. Though these particles are not infectious, it is known that particles
larger than 5 m can damage or even block blood vessels leading to severe adverse effects.
Particle contamination can even lead to embolisms in the brain and respiratory system which can
be fatal.

What are the Pharmacopoeial requirements?

Solutions for injection administered by the intramuscular or subcutaneous route must meet the
requirement of particle matter in injection. Parenteral preparation including solution constituted
from sterile solids are expected to be free from particles of approximately 50m or more that can
be observed by inspection with the unaided eye. However, Parenteral preparations in containing
100 ml or more of a single dose large volume injection intended for administration by
intravenous infusion should comply with the limits of sub visible particle prescribed in this test.

The test does not apply to multiple dose injections, to single dose small volume parenteral
preparations and to parenteral solutions constituted from sterile solids.
What are the prescribed methods?
1. Light Obscuration Particle Count Test
2. Microscopic Particle Count Test
3. Visual inspection

What is the principle behind the light obscuration liquid borne particle counter?
The liquid particle counter operates on the principle that the light extinguished by a particle in a
liquid within a laser beam is direct function of its area. Particles obscure the laser beam during
transit through the laser beam. The pulses produced by electronically detecting the total laser
light minus the light obscured by the particle are used to size the particle. These pulses are
measured by an analog to digital converter in the sensor. The liquid is presented to the optical
system through a rectangular capillary. The capillary has a window attached to both the front and
back sides which are coated to reduce reflections. A central software application controls the
hardware, analyzes the data and stores the data future interpretation. The variation in light caused
by passing of a particle is electronically detected by the photodetecter. This signal is than
amplified and converted to its digital equivalent. The value of this digital signal is converted into
an equivalent particle size in a microprocessor. The different size particles are counted and stored
and made available for transmission to the data display system upon request.

Schematic diagram of Light Obscuration Based Liquid Particle Counter

The test is carried out under conditions limiting particulate matter, preferably in a laminar-flow
cabinet.
What are the applications for the liquid borne particle counter?
Despite the most obvious application of pharmaceutical parenteral monitoring to USP, EP, JP,
and FDA standards, the system can also be used for the following
1. Parts/medical device cleanliness testing
2. Laboratory water sampling
3. WFI and Purified Water testing
4. Filter efficiency testing

Cryoscopic Osmometer

Osmometer Based on Freezing Point Depression

Osmolality measurements using a osmometer, are regularly carried out in the pharmaceutical
industry and research labs to establish the isotonicity of intravenous solutions, injection, nasal
and eye drops. Preparations which are hypertonic or hypotonic can cause severe irritation when
injected into the bloodstream or tissue or when applied to mucous membranes like cornea and
wounds.
Normal ranges of osmolality
The osmolality of blood, measured using osmometer, ranges from 250 to 350 mOsm/kg and the
normal osmolarity of body fluids given in medical books ranges from 275 to 295 mOsm/kg, but
normal values usually fall in an even narrower range of 2864 mOsm/kg. Parenteral preparations
are developed to closely match the values to reduce the irritation caused by them
Pharmacopoeia Requirement
Osmolality measurement using osmometer is a pharmacopoeia requirement for numerous
products. The United States Pharmacopoeia (USP) gives the option of using either vapour

pressure based osmometer or a cryoscopic (depression increasing point based) osmometer for the
measurement of osmolarity. Whereas the Indian Pharmacopoeia (IP) only describes the
cryoscopic osmometer for the measurement of osmolarity.
Features of osmometer
The system has numerous advantages such as

Automated calibration

A short measurement time of 1 min

Automated measurement and calculation

Low sample volume of only 50 l

Built-in printer and

Three point calibration

Reproducibility of less than 0.5% for 50 m samples

A large measuring range of up to 3000 mOsm/kg

A resolution of 1 mOsm/kg over the entire measuring range

The system has been installed and duly qualified as per our internal quality systems which are
accredited by NABL to ISO/IEC 17025 and is now ready for service for our customers in various
industries.
Introduction to basic principles of osmometry
Osmosis is defined as the passage of solvent into a solution through a semi permeable
membrane. This process eventually leads to equilibrium and the tendency of the solvent to move

from one side of the membrane to the other can be measured by the closely related colligative
property called osmotic pressure.
Osmotic pressure is classically given in terms of atmospheres, but in clinical practice, it is
expressed in terms of osmols (Osm) or milliosmols (mOsm). 1-osmolal solution is defined as the
solution containing one mole of a non-ionising substance in 1 kg of water. It is a measure of the
total number of particles dissolve in a kilogram of water and depends on the electrolytic nature of
the solute, an ionic solute will dissociate in water to form ions or particles. The apparent number
of particles in solution is measured using an osmometer.
Osmolarity is used more frequently than osmolality in the labelling of parenteral solutions in,
pharmaceutical manufacturing, hospitals and pharmacies. But osmolarity cannot be measured
and must be calculated experimentally by data mining the osmolality of a solution.
For near ideal solutions, osmolarity different from osmolality only by 1 or 2% , but as you go
towards more concentrated solutions of polyvalent electrolyte together with buffers,
preservatives and other ions these differences become larger.
For accuracy in the preparation and labelling of parenteral solutions osmolality should be
measured carefully with the vapour pressure or freezing point based osmometer rather than being
calculated based on the concentrations of ingredients added.

Water Activity Meter

Water activity is an important parameter employed in definition of stability of foods. The water
activity should lie within defined limits for a desired shelf life of food products. The value should
be below the limit to prevent mould growth. In addition ,undesirable reactions such as nonenzymatic browning, fat oxidation vitamin degradation, enzymatic reactions, protein
denaturation, starch geltinization and starch retrogradation are also slowed below the specified
water activity .Measurements can be made conveniently below 5 minutes using the Water
Activity Meter. It is a PC controlled system based on capacitance measuments run on HW4
software. Simultaneous loading of upto 4 samples is possible in a single run.